Sphingolipids and acylcarnitines are altered in placentas from women with gestational diabetes mellitus.

Gestational diabetes mellitus (GDM) is the most common medical complication of pregnancy and a severe threat to pregnant people and offspring health. The molecular origins of GDM, and in particular the placental responses, are not fully known. The present study aimed to perform a comprehensive characterisation of the lipid species in placentas from pregnancies complicated with GDM using high-resolution MS lipidomics, with a particular focus on sphingolipids and acylcarnitines in a semi-targeted approach. The results indicated that despite no major disruption in lipid metabolism, placentas from GDM pregnancies showed significant alterations in sphingolipids, mostly lower abundance of total ceramides. Additionally, very long-chain ceramides and sphingomyelins with twenty-four carbons were lower, and glucosylceramides with sixteen carbons were higher in placentas from GDM pregnancies. Semi-targeted lipidomics revealed the strong impact of GDM on the placental acylcarnitine profile, particularly lower contents of medium and long-chain fatty-acyl carnitine species. The lower contents of sphingolipids may affect the secretory function of the placenta, and lower contents of long-chain fatty acylcarnitines is suggestive of mitochondrial dysfunction. These alterations in placental lipid metabolism may have consequences for fetal growth and development.

Jussara berry (Euterpe edulis M.) oil-in-water emulsions are highly stable: the role of natural antioxidants in the fruit oil.

BACKGROUND: Antioxidants help prevent lipid oxidation, and therefore are critical to maintain sensory quality and chemical characteristics of edible oils. Jussara berry (Euterpe edulis M.) oil is a source of minor compounds with potential antioxidant activity. The aim of this work was to investigate the role of such compounds on the effectiveness to prevent or delay oxidation of oil present in oil-in-water emulsions, and how the emulsions' physical stability would be affected. RESULTS: Jussara berry oil extracted by ethanol extraction, its stripped variations (partially stripped, highly stripped and highly stripped with added butylhydroxytoluene), and expeller-pressed oil were used to prepare oil-in-water emulsions. Jussara berry oils were analyzed before emulsions preparation to ensure their initial quality and composition, and oil-in-water emulsions were analyzed regarding their oxidative and physical stability. Ethanol extracted oil emulsion presented higher oxidative stability than highly stripped oil emulsion with added synthetic antioxidant butylahydroxytoluene (oxidative stability index 45% lower, after 60 days, and reached undetectable levels after 90 days). All emulsions remained physically stable for up to 120 days of storage. CONCLUSION: Our results indicate that natural antioxidants in jussara berry oil protect emulsions from oxidation while keeping physical stability unchanged. © 2018 Society of Chemical Industry.

Use of Yarrowia lipolytica Lipase Immobilized in Cell Debris for the Production of Lipolyzed Milk Fat (LMF).

Lipase immobilized on Yarrowia lipolytica cell debris after sonication of yeast cells (LipImDebri) was used in hydrolysis reaction as a novel strategy to produce lipolyzed milk fat (LMF). Extracellular (4732.1 U/L), intracellular (130.0 U/g), and cell debris (181.0 U/g) lipases were obtained in a 4 L bioreactor using residual frying oil as inducer in 24 h fermentation process. LipImDebri showed a good operational stability retaining 70% of lipolytic activity after the second cycle and 40% after the fourth. The highest degree of hydrolysis (28%) was obtained with 500 mg LipImDebri for 6 h of lipolysis of anhydrous milk fat. LMF produced with LipImDebri presented high contents of oleic (35.2%), palmitic (25.0%), and stearic (15.4%) acids and considerable amounts of odor-active short and medium chain fatty acids (C:4⁻C:10) (8.13%).

Anthocyanin-Rich Grape Pomace Extract (Vitis vinifera L.) from Wine Industry Affects Mitochondrial Bioenergetics and Glucose Metabolism in Human Hepatocarcinoma HepG2 Cells.

Cancer cells demand high ATP provisions to support proliferation, and targeting of energy metabolism is a good strategy to increase their sensitivity to treatments. In Brazil, wine manufacture is expanding, increasing the amount of pomace that is produced. We determined the phenolic composition and antioxidant properties of a dark skin Grape Pomace Extract and its effects on metabolism and redox state in human hepatocarcinoma HepG2 cells. The material and the methods used represented the industrial process since pomace derived from white wine production and the extract concentrated by pilot plant scale reverse osmosis. Grape pomace extract was rich in polyphenols, mainly anthocyanins, and presented high antioxidant capacity. Short-term metabolic effects, irrespective of any cytotoxicity, involved increased mitochondrial respiration and antioxidant capacity and decreased glycolytic metabolism. Long-term incubation was cytotoxic and cells died by necrosis and GPE was not toxic to non-cancer human fibroblasts. To the best of our knowledge, this is the first report to characterize pomace extract from white wine production from Brazilian winemaking regarding its effects on energy metabolism, suggesting its potential use for pharmaceutical and nutraceutical purposes.

High hydrostatic pressure processing affects the phenolic profile, preserves sensory attributes and ensures microbial quality of jabuticaba (Myrciaria jaboticaba) juice.

BACKGROUND: Jabuticaba (Myrciaria jaboticaba) is a Brazilian fruit rich in phenolic compounds and much appreciated for its sweet and slightly tangy taste. However, the high perishability of this fruit impairs its economic exploitation, creating an opportunity for the development of innovative products, such as high hydrostatic pressure (HHP) processed juices. We investigated the effect of HHP (200, 350 and 500 MPa for 5, 7.5 and 10 min) on phenolic compounds, antioxidant activity and microbiological quality of jabuticaba juice and the effect of the most effective HHP condition on its sensory acceptance. RESULTS: Pressurization increased total phenolic compound content (up to 38%) and antioxidant activity by FRAP assay (up to 46%), probably by increasing phenolic compound extractability due to tissue damage. Pressurization progressively decreased microbial counts, and colony growth was undetectable at pressures of 350 MPa or 500 MPa. With the exception of aroma, which was 10% lower in pressurized juice at 350 MPa for 7.5 min in relation to unprocessed juice, HHP did not affect sensory acceptance scores. CONCLUSION: Our results show that HHP was effective in ensuring microbiological quality, increasing bioactive potential and maintaining overall acceptance of jabuticaba juice, reinforcing the potential application of this processing technology in bioactive-rich foods. © 2017 Society of Chemical Industry.

Optimized extraction of polyphenolic antioxidant compounds from Brazil nut (Bertholletia excelsa) cake and evaluation of the polyphenol profile by HPLC.

BACKGROUND: The solid residue (cake) of pressed Brazil nut oil has high energy value and contains high levels of nutrients and bioactive compounds, such as polyphenols. However, little is known about these components in this by-product. Extraction is the first step in investigating the phenolic compounds in Brazil nut cake because extraction conditions might impact the yields of phenolic compounds and antioxidant capacity. The aim of this study was to select the best phenolic compound extraction conditions for Brazil nut cake by using factorial experimental design and to characterize the phenolic compounds in the extract. RESULTS: The optimal extraction of antioxidant phenolic compounds from Brazil nut cake was achieved under the following conditions: ethanol-water (40:60; v/v); 2.5 min homogenization; and 1 h extraction at 60 °C. The phenolic compound profile of the Brazil nut cake extract using the optimized extraction was determined by high-performance liquid chromatography with photodiode array detection. Six phenolic acids (gallic acid, protocatechuic acid, 2,4-dihydroxybenzoic acid, p-hydroxybenzoic acid, p-coumaric acid and sinapic acid) and one flavonoid ((+)-catechin) were identified, and the contents of the phenolic compounds varied from 70.0 to 421 mg kg(-1) . CONCLUSION: Knowledge of the potential bioactivity of Brazil nut cake identified in the present study might promote its use in the food industry. © 2015 Society of Chemical Industry.

Bread formulated with guava powder was enriched in phenolic and aroma compounds, and was highly acceptable by consumers.

Guava powder (GP) was used as source of aroma and phenolic compounds to fortify wheat bread 10% (GB10) and 20% (GB20), substituting for wheat flour. Phenolic compounds, antioxidant capacity, volatile compounds profile, and sensory acceptability of control bread (CB; without GP) and guava breads (GB) were evaluated. Incorporation of GP increased roughly 2-to-3-fold the phenolic compounds contents of bread. Ten phenolic compounds were identified in GB20, and quercetin-3-O-rutinoside was the major compound, while in CB, ferulic acid was the major among the six phenolic compounds in CB. Bread making seemed to promote the release of phenolic compounds from structural components. Breads incorporated with GP presented a richer volatile profile than CB, especially due to the presence of terpenes. GB improved aroma profile of bread. GP added aroma compounds and phenolic antioxidants, and seemed to be an interesting approach to enhance bread bioactivity and acceptability.

Placental F4-Neuroprostanes and F2-Isoprostanes are altered in gestational diabetes mellitus and maternal obesity.

We investigated whether gestational diabetes mellitus (GDM) associated with maternal obesity modifies the placental profile of F4-Neuroprostanes and F2-Isoprostanes, metabolites of non-enzymatic oxidation of docosahexaenoic acid (DHA) and arachidonic acid (AA), respectively. Twenty-five placental samples were divided into lean (n=11), obesity (n=7) and overweight/obesity+GDM (n=7) groups. F4-Neuroprostanes and F2-Isoprostanes were higher in obesity compared to lean controls, but reduced to levels similar to lean women when obesity is further complicated with GDM. Lower content of F2-Isoprostanes suggests adaptive placental responses in GDM attenuating oxidative stress. However, low levels of placental F4-Neuroprostanes may indicate impaired DHA metabolism in GDM, affecting fetal development and offspring health. These results were not related to differences in placental content of DHA, AA and polyunsaturated fatty acids status nor to maternal diet or gestational weight gain. Placental DHA and AA metabolism differs in obesity and GDM, highlighting the importance of investigating the signalling roles of F4-Neuroprostanes and F2-Isoprostanes in the human term placenta.

Integrated NMR and MS Analysis of the Plasma Metabolome Reveals Major Changes in One-Carbon, Lipid, and Amino Acid Metabolism in Severe and Fatal Cases of COVID-19.

Brazil has the second-highest COVID-19 death rate worldwide, and Rio de Janeiro is among the states with the highest rate in the country. Although vaccine coverage has been achieved, it is anticipated that COVID-19 will transition into an endemic disease. It is concerning that the molecular mechanisms underlying clinical evolution from mild to severe disease, as well as the mechanisms leading to long COVID-19, are not yet fully understood. NMR and MS-based metabolomics were used to identify metabolites associated with COVID-19 pathophysiology and disease outcome. Severe COVID-19 cases (n = 35) were enrolled in two reference centers in Rio de Janeiro within 72 h of ICU admission, alongside 12 non-infected control subjects. COVID-19 patients were grouped into survivors (n = 18) and non-survivors (n = 17). Choline-related metabolites, serine, glycine, and betaine, were reduced in severe COVID-19, indicating dysregulation in methyl donors. Non-survivors had higher levels of creatine/creatinine, 4-hydroxyproline, gluconic acid, and N-acetylserine, indicating liver and kidney dysfunction. Several changes were greater in women; thus, patients' sex should be considered in pandemic surveillance to achieve better disease stratification and improve outcomes. These metabolic alterations may be useful to monitor organ (dys) function and to understand the pathophysiology of acute and possibly post-acute COVID-19 syndromes.

Pomegranate (Punica granatum) peel fractions obtained by supercritical CO2 increase oxidative and colour stability of bluefish (Pomatomus saltatrix) patties treated by UV-C irradiation.

The sequential fractionation by supercritical-CO2 (SC-CO2) was applied to obtain fractions enriched in bioactive compounds of pomegranate peel, and we investigated if pomegranate peel extract and fractions would be effective to inhibit lipid and protein oxidation, and discolouration of bluefish patties stored at 4 °C for 9 days, after UV-C irradiation. The non-fractionated SC-CO2 extract from pomegranate peel was rich in phenolic compounds, mainly ellagitannins, besides, it possessed lipophilic compounds such as tocopherols and ß-carotene. These compounds were successfully separated by the fractionation protocols, in a lipid fraction concentrated in lipophilic compounds, and one or two fractions enriched with phenolic compounds, especially ellagitannins. The lipid fraction and the high phenolics fraction from pomegranate peel were then as effective as the synthetic antioxidant BHT in avoiding bluefish patties oxidation during refrigerated storage. Our data indicates that pomegranate peel fractions could be used to replace a synthetic antioxidant in fish meat.

Palm oil wastes as feedstock for lipase production by Yarrowia lipolytica and biocatalyst application/reuse.

Palm oil production chain generates a greasy residue in the refining stage, the Palm Oil Deodorizer Distillate (PODD), mainly composed of free fatty acids. Palm oil is also used industrially to fry foods, generating a residual frying oil (RFO). In this paper, we aimed to produce lipase from palm agro-industrial wastes using an unconventional yeast. RFO_palm, from a known source, consisted of 0.11% MAG + FFA, 1.5% DAG, and 97.5 TAG, while RFO_commercial, from a commercial restaurant, contained 6.7% of DAG and 93.3% of TAG. All palm oil wastes were useful for extracellular lipase production, especially RFO_commercial that provided the highest activity (4.9 U/mL) and productivity (465 U/L.h) in 75 h of processing time. In 48 h of process, PODD presented 2.3 U/mL of lipase activity and 48.5 U/L.h of productivity. RFO_commercial also showed the highest values for lipase associated to cell debris (843 U/g). This naturally immobilized biocatalyst was tested on hydrolysis reactions to produce Lipolyzed Milk Fat and was quite efficient, with a hydrolysis yield of 13.1% and 3-cycle reuse. Therefore, oily palm residues seem a promising alternative to produce lipases by the non-pathogenic yeast Y. lipolytica and show great potential for industrial applications.

Decreased Fatty Acid Transporter FABP1 and Increased Isoprostanes and Neuroprostanes in the Human Term Placenta: Implications for Inflammation and Birth Weight in Maternal Pre-Gestational Obesity.

The rise in prevalence of obesity in women of reproductive age in developed and developing countries might propagate intergenerational cycles of detrimental effects on metabolic health. Placental lipid metabolism is disrupted by maternal obesity, which possibly affects the life-long health of the offspring. Here, we investigated placental lipid metabolism in women with pre-gestational obesity as a sole pregnancy complication and compared it to placental responses of lean women. Open profile and targeted lipidomics were used to assess placental lipids and oxidised products of docosahexaenoic (DHA) and arachidonic acid (AA), respectively, neuroprostanes and isoprostanes. Despite no overall signs of lipid accumulation, DHA and AA levels in placentas from obese women were, respectively, 2.2 and 2.5 times higher than those from lean women. Additionally, a 2-fold increase in DHA-derived neuroprostanes and a 1.7-fold increase in AA-derived isoprostanes were seen in the obese group. These changes correlated with a 70% decrease in placental FABP1 protein. Multivariate analyses suggested that neuroprostanes and isoprostanes are associated with maternal and placental inflammation and with birth weight. These results might shed light on the molecular mechanisms associated with altered placental fatty acid metabolism in maternal pre-gestational obesity, placing these oxidised fatty acids as novel mediators of placental function.

Efficient production of bioactive structured lipids by fast acidolysis catalyzed by Yarrowia lipolytica lipase, free and immobilized in chitosan-alginate beads, in solvent-free medium.

Structured lipids (SL) represent a new generation of lipids, considered bioactive compounds. Medium-chain, oleic (18:1n-9), and medium-chain fatty acid (MCFA) structured lipids (MOM-SL) were produced by acidolysis reaction in solvent-free medium with capric (10:0) and lauric (12:0) free fatty acids (FFAs) and triolein or olive oil, using Yarrowia lipolytica lipase as biocatalyst. MCFAs were rapidly incorporated into sn-1,3 SL in acidolysis reactions with triolein and olive oil, up until 30% of incorporation efficiency of capric and lauric acids in SLs. The kinetics of MCFA incorporation in MOM-SL was influenced by the FFA:TAG molar ratio, and for reactions between triolein and lauric acid, increasing FFA:TAG from 2:1 to 4:1 enhanced MCFA incorporation in SL. Y. lipolytica lipase showed a strictly 1,3-regioselective profile in acidolysis reaction, confirmed by nuclear magnetic resonance spectroscopy. Immobilization of this lipase by microencapsulation in chitosan-alginate beads resulted in similar incorporation efficiency for lauric acid with olive oil TAG and this reaction could be performed for 5 cycles without catalytic activity loss. This lipase showed promising properties as a potential biocatalyst that may be effectively used in production of bioactive structured lipids, which might be applied for prevention of metabolic and inflammatory disorders related to obesity.

Erythrocyte membrane and plasma non-esterified n-3 and n-6 polyunsaturated fatty acids of pregnant and non-pregnant Brazilian adolescents.

The aim of the present study was to evaluate the influence of pregnancy in adolescents on the fatty acid composition of the erythrocyte membrane, which was used as a proxy for status of n-3 and n-6 polyunsaturated fatty acids (PUFA), and also on the composition of plasma non-esterified fatty acids (NEFA) mobilized from the adipose tissue. Two matched groups of healthy adolescents (14-19 years) from Rio de Janeiro, Brazil, were compared: pregnant (n=26; 32.7+/-3.9 weeks of gestation, mean+/-SD) and non-pregnant (n=20). Blood samples were collected after an overnight fast. Mean dietary intakes of total fat and n-3 and n-6 PUFA (energy %) were not different between pregnant and non-pregnant adolescents, and the consumption of food sources of docosahexaenoic acid (DHA) was low. Fasting total NEFA and NEFA 18:2n-6, 18:3n-6 and 20:4n-6 (g/100g fatty acids) were higher in pregnant than in non-pregnant adolescents. Although erythrocyte 20:4n-6 was lower in pregnant adolescents, there were no differences in DHA (g/100g fatty acids), in DHA status indices (22:5n-6/22:4n-6 and 22:6n-3/22:5n-6 ratios) and in the index of n-3+n-6 PUFA status ([Sigman-3+Sigman-6]/[Sigman-7+Sigman-9]) in erythrocytes as compared with those of non-pregnant adolescents. In conclusion, pregnancy did not have an adverse effect on erythrocyte DHA content or on DHA and n-3+n-6 PUFA status indices in the adolescents studied.

Fermentation of Milk into Yoghurt and Cheese Leads to Contrasting Lipid and Glyceride Profiles.

There is mounting evidence that the consumption of fermented dairy products such as cheese and yoghurt is associated with a reduced risk of type II diabetes. This effect is greater than in fresh milk and differs between cheese and yoghurt. However, the molecular components responsible for the effect are not known. We tested the hypothesis that the lipid and/or glyceride profiles of yoghurts and cheeses are distinct from one another and fresh milk. We developed a novel sample preparation technique for high-fat samples that can be used with Direct Infusion-Mass Spectrometry. We found that the lipid and glyceride profiles of cheddars from the UK, Ireland and France, and hard cheeses from Sweden and Italy were similar to one another but distinct from unfermented dairy products. The lipid and glyceride profile of yoghurts was varied and included types that may be similar to fresh milk. Several odd-chain-containing triglycerides were more abundant, while a variety of others were less abundant, in fermented milk samples. Phosphatidylcholines and phosphatidylethanolamines were more abundant in cheeses, with evidence that the phosphatidylethanomine profile is re-modelled in a way that reflects the bacterial cell envelope. We concluded that a combination of microorganismal metabolism, concentration of the lipid/glyceride fraction and oxidation during fermentation contribute to the observed lipid profile if fermented dairy foods. These differences in the lipid and glyceride profile provide a new avenue for understanding why different fermented dairy foods show a different association with reduced disease risk compared to unfermented dairy.

Breast Milk Content of Vitamin A and E from Early- to Mid-Lactation Is Affected by Inadequate Dietary Intake in Brazilian Adult Women.

Our aims were to investigate vitamin A and E status during lactation and the determinants of breast milk content for the appropriate nutrition of the infant in a study with nursing Brazilian women. We hypothesized that both inadequate intake and the lipoprotein distribution of vitamin A and E during lactation could have an impact on their breast milk levels from early- to mid-lactation. Nineteen adult lactating women participated in this longitudinal observational study, in which dietary records, blood and mature breast milk samples were collected for the analysis of vitamin A and E, and carotenoids in early- (2nd to 4th week) and mid-lactation (12th to 14th week). Nutrient intake was balanced by the Multiple Source Method (MSM), and the intake of vitamin A and E was inadequate in 74 and 100% of the women, respectively. However, these results were not reflected in low serum concentrations of retinol and only 37% of the volunteers were vitamin E deficient according to the blood biomarker. As lactation progressed, vitamin A and E status worsened, and this was clearly observed by the decrease in their content in breast milk. The reduced content of vitamin A and E in the breast milk was not related to their distribution in lipoproteins. Taken together, the contents of vitamin A and E in breast milk seemed to be more sensitive markers of maternal nutrition status than respective blood concentrations, and dietary assessment by the MSM in early lactation was sensitive to indicate later risks of deficiency and should support maternal dietary guidance to improve the infant's nutrition.

Essential and long-chain polyunsaturated fatty acid status and fatty acid composition of breast milk of lactating adolescents.

The aims of the present study were to evaluate essential fatty acids (EFA) and long-chain PUFA (LCPUFA) status in lactating adolescents and its association with breast milk composition. Healthy nursing adolescents from Rio de Janeiro, Brazil (n 30; 14-19 years; 30-120 d postpartum), exclusively or predominantly breast-feeding, participated in this study. Breast milk and blood samples were collected after overnight fasting. Fatty acid composition of breast milk, erythrocyte membrane (EM) and plasma NEFA were determined by GC. Indices of fatty acid status (mean melting point (MMP); EFA status index; DHA status indices, 22 : 5n-6:22 : 4n-6 and 22 : 6n-3:22 : 5n-6 ratios) were calculated from EM fatty acid composition. Dietary intake of n-3 fatty acids was low when compared with current recommendations for lactating women. MMP was associated with indices of DHA status, some individual fatty acids in EM and years post-menarche and weeks postpartum, suggesting the use of erythrocyte MMP as a possible comprehensive biochemical marker of LCPUFA status in this physiological condition. The DHA status of lactating adolescents and their milk DHA concentrations were similar to the values of Brazilian lactating adults, but lower compared with the values of lactating adults from other countries. Therefore, these lactating adolescents were apparently not disadvantaged, as compared with the Brazilian adults, when EM and breast milk fatty acid composition were considered. In general, PUFA in milk from adolescents presented few associations with their concentrations in plasma NEFA and with maternal status. However, milk DHA was associated with maternal LCPUFA and DHA states.

Indices of dietary fat quality during midpregnancy is associated with gestational diabetes.

OBJECTIVE: To investigate the relationship between gestational diabetes mellitus (GDM) and the usual intake of fatty acids and indices of dietary fat quality [the atherogenicity (AI) and thrombogenicity indices (TI), and the ratios of hypo-and hypercholesterolemic (hH), ∑n-3/∑n-6, and polyunsaturated/saturated fatty acids (P:S)], during mid-pregnancy. METHODS: 799 adult pregnant women living in Ribeirão Preto, SP, Brazil were screened and accepted for this cross-sectional GDM study. The Multiple Source Method was used to estimate participants' usual diet, using two 24-hour dietary recalls during mid-pregnancy. Diagnosis of GDM was defined by the American Diabetes Association criteria of 2015. Logistic regression analysis were used to assess the association between GDM and dietary fat, adjusted for age, education, parity, gestational age at the time of the interview, pre-pregnancy and current BMI, prior GDM, family history of diabetes, smoking, physical activity, energy, fiber, and fatty acids. RESULTS: The mean (standard deviation) age of the women was 28 (5) years, and 19% had GDM. After multiple adjustments, inverse associations between the highest tertile of ∑n-3 fatty acids intake [0.21 (0.08-0.56)], α-linolenic intake [0.15 (0.05-0.42)], and GDM were found. A positive association between GDM and the highest tertile of TI [2.66 (1.34-5.29)], and a negative association with the highest tertile of hH ratio [0.41 (0.22-0.77)], were observed. No association between GDM and other indices of dietary fat quality were found. CONCLUSION: The quality of dietary fat during pregnancy is strongly associated with GDM. Randomized clinical trial are needed to confirm these findings.

Nutrient composition of banked human milk in Brazil and influence of processing on zinc distribution in milk fractions.

OBJECTIVE AND METHODS: We measured the contents of fat, protein, lactose, calcium, phosphorus, zinc, iron, copper, and vitamin A in processed mature milk samples (individual, n = 60, and pooled, n = 10) from a reference human milk bank in Brazil and assessed the effect of pasteurization followed by freezing on the nutrient composition and the pattern of zinc distribution in fractions (fat, whey, and casein) of milk samples (n = 15). RESULTS: Mean nutrient concentrations were within expected ranges in mature milk from healthy women, except fat, which was lower. Interindividual variability of nutrient concentrations was high (coefficient of variation, 21-62%) but reduced overall in pooled samples. Processing of milk samples did not affect the nutrient contents but did cause a significant shift (P < 0.04) in the relative distribution of zinc, with a decrease in the whey fraction and an increase in the fat fraction. CONCLUSIONS: Redistribution and possible alterations in the zinc-binding pattern during processing in human milk banks may reduce zinc bioavailability to the infant.

Mathematical method for the prediction of retention times of fatty acid methyl esters in temperature-programmed capillary gas chromatography.

An accurate method for identification of fatty acids in complex mixtures analyzed by temperature-programmed capillary gas chromatography is described. The method is based on a mathematical approach using regression curves obtained by plotting the relative retention times of fatty acid methyl esters (FAMEs) analyzed in isothermal and gradient temperature conditions. The method was applied to a complex biological sample (human milk), and it was possible to identify 64 fatty acids, including branched-chain and other fatty acids for which reference standards were not readily available. The identities of the majority of the peaks were confirmed by mass spectrometry. The relative residuals and the relative differences between estimated and measured relative retention times of individual FAMEs varied from 0.03 to 3.15% and from 0.0 to 2.9%, respectively. The method is useful for identification of fatty acids in routine analysis.